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  • DNase I, from Bovine Pancreas
  • 產品型號:101-9003-98-9

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Used for the removal of DNA from protein samples. Synonym: Deoxyribonuclease I, Deoxyribonucleate 5′-oligonucleotido-hydrolase

Appearance: white powder, occasionally off-white or with light yellow cast.
Activity: >300 Kunitz U/mg
Residue on ignition: <1%

Highly purified by chromatography to remove trace contaminating RNase and proreases.

Deoxyribonuclease I (DNase I) is an endonuclease that hydrolyzes double-stranded or single-stranded DNA preferentially at sites adjacent to pyrimidine nucleotides. The product of hydrolysis is a complex mixture of 5'-phosphate mononucleotides and oligonucleotides. In the presence of magnesium ion, DNase I attacks each strand of DNA independently and the cleavage sites are random. In the presence of manganese (II), DNase I cleaves both strands of DNA at approximately the same site. Most protocols use magnesium ion with DNase I but for specific purposes, manganese is cited.

Preparation Instructions:
For long term storage, reconstitute with water or any buffer, pH 4.0–9.0, except phosphate buffer and avoid calcium chelators. Addition of 50% glycerol will maintain a liquid state at -20 °C without affecting stability.
Only freeze and thaw solutions once.

Storage/Stability:
This lyophilized DNase I product retains activity for 2-3 years when stored at 2-8 °C. Solutions of DNase I (10 mg/ml) in 0.15 M NaCl may lose <10% of its activity stored for a week in aliquots at -20 °C. The same solutions stored in aliquots at 2–8 °C can lose ~20% activity.

DNase I remains active in solution between pH 5 and 7 up to 60 °C for at least five hours. A 1 mg/ml solution in acetate buffer (pH 5.0) or Tris buffer (pH 7.2) loses activity at the rate of 6%/hour. At 68 °C DNase I loses activity in <10 minutes.

Usage:
DNase I is used to remove DNA from protein and nucleic acid samples, and to nick DNA as a first step to incorporate labeled bases into DNA.
For complete digestion of DNA, 1 mg of DNA can be digested with 1–2 units of DNAse at 25–37 °C for 10 minutes in 40 mM Tris-HCl, 100 mM NaCl, 60 mM MgCl2, and 10 mM CaCl2, pH 7.9.
Store at -20ºC.